Molecular cloning and expression of cDNA encoding a lumenal calcium binding glycoprotein from sarcoplasmic reticulum.
Molecular cloning and expression of cDNA encoding a lumenal calcium binding glycoprotein from sarcoplasmic reticulum.
复制标题
编码来自肌浆网的腔内钙结合糖蛋白的 cDNA 的分子克隆和表达。
DOI：
10.1073/pnas.86.16.6047
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发表时间：
1989
影响因子：
11.1
通讯作者：
D. Maclennan
中科院分区：
文献类型：
--
作者：
E. Leberer;Jeffrey H. M. CHARUKt;N. M. GREENt;D. Maclennan
用抗体筛选的方法分离到编码兔骨骼肌肌浆网160 kDa糖蛋白的基因。该基因与编码53 kDa糖蛋白的基因完全相同，只是在其5‘端含有1308个核苷酸的框内插入，显然是由选择性剪接引起的。该基因编码的蛋白含有一个19个残基的NH2末端信号序列和一个与53 kDa糖蛋白相同的453个残基的COOH末端序列。它还将在这些序列之间插入一个436个氨基酸。这个插入物将是高度酸性的，表明它可能结合了钙离子。凝胶覆盖实验表明，纯化的160 kDa糖蛋白和在COS-1细胞中表达的160 kDa糖蛋白可与45Ca+结合。该蛋白定位于肌浆网的管腔内，并通过钙离子与肌浆网膜结合。我们建议将肌浆网的这种腔内钙结合糖蛋白命名为“Sarcalumenin”。
Antibody screening was used to isolate a cDNA encoding the 160-kDa glycoprotein of rabbit skeletal muscle sarcoplasmic reticulum. The cDNA is identical to that encoding the 53-kDa glycoprotein except that it contains an in-frame insertion of 1308 nucleotides near its 5' end, apparently resulting from alternative splicing. The protein encoded by the cDNA would contain a 19-residue NH2-terminal signal sequence and a 453-residue COOH-terminal sequence identical to the 53-kDa glycoprotein. It would also contain a 436-amino acid insert between these sequences. This insert would be highly acidic, suggesting that it might bind Ca2+. The purified 160-kDa glycoprotein and the glycoprotein expressed in COS-1 cells transfected with cDNA encoding the 160-kDa glycoprotein were shown to bind 45Ca2+ in a gel overlay assay. The protein was shown to be located in the lumen of the sarcoplasmic reticulum and to be associated through Ca2+ with the membrane. We propose that this lumenal Ca2+ binding glycoprotein of the sarcoplasmic reticulum be designated "sarcalumenin."